High efficiency transformation with lithium acetate
1992; Oxford University Press; Linguagem: Inglês
10.1093/oso/9780199634309.003.0008
AutoresR. Daniel Gietz, Robin A. Woods,
Tópico(s)Fungal and yeast genetics research
ResumoAbstract The transformation of lithium acetate (LiAc) treated yeast cells with plasmid DNA was first reported in 1983 (1). The procedure was less efficient than transformation of cells converted to sphaeroplasts by treatment with Zymolyase (2, 3), but was simpler and faster. The original procedure, and subsequent improvements reported by a number of workers, involved a pre-incubation of the cells in LiAc, followed by an incubation in LiAc plus plasmid DNA, then by the addition of polyethylene glycol (PEG), and finally by exposure to a heat shock. The yields of transformants with this procedure rarely exceeded 1 × 10/µg plasmid DNA (4), compared with 2 × 105/µg for the sphaeroplast procedure (5). In 1989 it was reported that inclusion of single-stranded carrier nucleic acids (DNA or RNA) in the transformation reaction increased the efficiency of the LiAc procedure dramatically; to 1 x 10 transformants/ µg (6). Further modifications to this ‘LiAc/ssDNA/PEG’ method reduced the time required and increased the efficiency more than 20 fold (7, 8). The current version of the method, described here, is markedly simpler and has yielded up to 2.2 x 107 transformants/µg plasmid DNA.
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